Application of Chromatographic Column Packed with Copolymer to Separation and Determination of Organic Acids by HPLC

The feasibility of application of copolymer packed chromatographic column using copolymer of polystryrene and divinylbenzene resin (PS-DVB) as packing material, to the separation of 5 standards of organic acids and 5 components in fermentation broth of succinic acid was studied. The PS-DVB based MKF-YJS column (250 mm×4.6 mm, 5 μm) with column temperature of 25 ℃ was used. The mobile phase containing 0.02 mol·L-1 NH4H2PO4 solution (adjust pH to 2.5 with H3PO4) and acetonitrile mixed in the ratio of 97∶3 (by vol.), with flow rate of 1.0 mL·min-1 was used. The detection wavelength was set at 210 nm. Five standard organic acids and five compounds in succinic acid fermentation broth were well separated by the proposed procedure. The calibration curve was in good linearity in the range of 0.05-12.0 g·L-1. The limit of detection was 0.005 g·L-1. The precision of sample introduction was 0.63% (n=5). The average recovery of succinic acid in fermentation broth was 91.8%.